Author: Stagegaard, Julia; Kurth, Andreas; Stern, Daniel; Dabrowski, Piotr Wojciech; Pocknell, Ann; Nitsche, Andreas; Schrick, Livia
Title: Seasonal recurrence of cowpox virus outbreaks in captive cheetahs (Acinonyx jubatus) Document date: 2017_11_9
ID: 0oa4x05s_15
Snippet: For ELISA, PolySorp microwell plates (Nunc) were coated with 100 μL of lysate of infected or non-infected HEp-2 cells at 4 μg/mL in 0.1 M carbonate buffer (pH 9.6) over night at 4˚C. Between each step, plates were washed with 4 × 300 μL of washing buffer (100 mM Tris, pH 7.5, 150 mM NaCl, 0.05% Tween 20). Plates were blocked at room temperature for 1 hour with 200 μL per well of 3% bovine serum albumin (BSA, Carl Roth) in washing buffer. Su.....
Document: For ELISA, PolySorp microwell plates (Nunc) were coated with 100 μL of lysate of infected or non-infected HEp-2 cells at 4 μg/mL in 0.1 M carbonate buffer (pH 9.6) over night at 4˚C. Between each step, plates were washed with 4 × 300 μL of washing buffer (100 mM Tris, pH 7.5, 150 mM NaCl, 0.05% Tween 20). Plates were blocked at room temperature for 1 hour with 200 μL per well of 3% bovine serum albumin (BSA, Carl Roth) in washing buffer. Subsequently, 100 μL per well of non-inactivated sera were incubated at a 1:100 dilution in washing buffer supplemented with 0.25% BSA for 1 hour before detection with either goat anti-cat IgG (H+L) HRP, goat anti-dog IgG (H+L) HRP, or goat anti-mouse IgG (H+L) (Dianova, used at a 1:5000 dilution). Finally, signals were developed for 15 minutes by using 100 μL of Seramun-Slow TMB substrate (Diavita) per well before the reaction was stopped by addition of 100 μL of 0.25 M H 2 SO 4 . The resulting absorption was read at 450 nm referenced to 620 nm at an ELISA reader (Tecan). Each serum was tested in two replicate measurements on both infected and non-infected HEp-2 cell lysate. Binding signals against non-infected HEp-2 cell lysate were subtracted from signals against vaccinia virus-infected cells, and sera with a differential binding signal above 0.05 were considered positive.
Search related documents:
Co phrase search for related documents- bovine serum albumin and carbonate buffer: 1, 2
- bovine serum albumin and cell lysate: 1, 2
- bovine serum albumin and ELISA reader: 1, 2, 3, 4
- bovine serum albumin and μl 100: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12
- bovine serum albumin and room temperature: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34
- bovine serum albumin and TMB substrate: 1, 2, 3, 4
- bovine serum albumin and vaccinia virus: 1, 2
- bovine serum albumin and wash buffer: 1, 2, 3
- bovine serum and BSA bovine serum albumin: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77
- bovine serum and bsa supplement: 1
- bovine serum and carbonate buffer: 1, 2, 3
- bovine serum and cell lysate: 1, 2, 3, 4, 5, 6
- bovine serum and ELISA reader: 1, 2, 3, 4
- bovine serum and μl 100: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33
- bovine serum and replicate measurement: 1
- bovine serum and room temperature: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53
- bovine serum and TMB substrate: 1, 2, 3, 4, 5, 6
- bovine serum and vaccinia virus: 1, 2, 3, 4, 5
- bovine serum and wash buffer: 1, 2, 3, 4, 5
Co phrase search for related documents, hyperlinks ordered by date